Acovenosigenin A -glucoside (AAG) is a cardiac glycoside derived from (Lour.) Merr, which exhibited the potential in treating lung cancer in our previous research. However, the action mechanism remains unclear. In this research, JAK2-STAT3 signaling pathway was predicted to be the critical regulation pathway based on the integrative analysis of transcriptome and proteome. Western blotting and qPCR assays were performed to identify that AAG can regulate JAK2-STAT3 signaling pathway and its downstream genes, such as c-Myc, Survivin, Cyclin B1, CDK1, Bcl-2. And this action of AAG depended on the suppression of STAT3 phosphorylation and its nuclear translocation through the experiments of Immunofluorescence, transient transfection and cryptotanshinone treatment. Additionally, AAG was discovered to mediate the JAK2-STAT3 pathway in IL-6-driven A549 and H460 cells, which in turn inhibited cell proliferation, promoted mitochondria-related apoptosis, and arrested the cell cycle progression. By molecular docking analysis, CETSA and SIP experiments, the protein of GP130 was identified as the specific target of AAG in A549 and H460 cells. Further studies suggested that AAG inhibited JAK2-STAT3 pathway and its downstream genes by targeting GP130 in nude mice xenograft model . This research presented that AAG exhibits the promising potential in the treatment of NSCLC.

中文翻译：

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基于转录组和蛋白质组综合分析及生物学验证，Acovenosigenin A β-葡萄糖苷通过靶向A549和H460细胞中的GP130介导JAK2-STAT3信号通路


Acovenosigenin A -glucoside (AAG) 是一种源自 (Lour.) Merr 的强心苷，在我们之前的研究中显示出治疗肺癌的潜力。然而，其作用机制仍不清楚。本研究基于转录组和蛋白质组的综合分析，预测JAK2-STAT3信号通路是关键的调控通路。通过Western blotting和qPCR检测发现AAG可以调节JAK2-STAT3信号通路及其下游基因，如c-Myc、Survivin、Cyclin B1、CDK1、Bcl-2。而AAG的这种作用依赖于通过免疫荧光、瞬时转染和隐丹参酮处理实验抑制STAT3磷酸化及其核转位。此外，还发现 AAG 在 IL-6 驱动的 A549 和 H460 细胞中介导 JAK2-STAT3 通路，进而抑制细胞增殖，促进线粒体相关细胞凋亡，并阻止细胞周期进程。通过分子对接分析、CETSA和SIP实验，确定GP130蛋白是A549和H460细胞中AAG的特异性靶点。进一步研究表明，AAG在裸鼠异种移植模型中通过靶向GP130抑制JAK2-STAT3通路及其下游基因。这项研究表明，AAG 在治疗 NSCLC 方面表现出巨大的潜力。